Protocols

• Breeding, Care And Maintenance Of Mice
• Purification Of BAC DNA For BAC Transgenics
• Detecting Transgenes by PCR and Southern Blot
• Preparing Earpunch DNA For PCR Genotyping
• Preparing Tail Or Toe DNA For Southern Blot Analysis
• Performing Gavage on Mice

David LePage's ES Cell Protocols:

LePage DF, Conlon RA. (2006) Animal models for disease: knockout, knock-in, and conditional mutant mice. Methods Mol Med 129:41-67. (.pdf)

• Growing ES Cells
• Passaging From 35 mm Dishes
• Freezing From 35 mm Dishes
• Preparing Vector DNA
• Electroporation
• Picking Lines
• Tryplating
• Freezing From 96-Well Plates
• Extracting Genomic DNA From 96-Well Plates
• Cutting Genomic DNA In 96-Well Plates
• Southerns
• Thawing 96-Well Plates
• Extracting Genomic DNA From 24-Well Plates Or Larger
• Counting ES Cell Chromosomes
• Counting ES Cell Chromosomes: Examples
• Preparing ES Cells For Blastocyst Injection

David LePage's Protocols for Making New ES Cell Lines:

pdf file

• References
• Schedule
• Initial Culture Conditions
• Embryo Collection
• Disaggregation
• Passaging
• Guidelines for Growing New Lines